Morning

Performed Calibration 1, 2, and 3 according to protocol. All measurements were performed using the plate reader of Seino.

Calibration 1

• L= 100 uL Ludox CL-X (stored at 4C)
• dd= 100 uL ddH20
• Measurement: Abs600, turn off pathlength correction

Calibration 2

• MS= 200 ul Microsphere Stock Solution
• dd= 100 uL ddH20
• green= serial dilution was performed with a micropipet from E1,F1,G1,H1 - E11,F11,G11,H11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
• Measurement: Abs600, re-mix befor putting in plate reader and prevent bubbles, path length correction off

Calibration 3

• 1xFC= 200 mL 1xFC (100uL 10x fluorescein + 900ul 1x PBS pH 7.4, tube was covered with foil
• P= 100 uL 1x PBS pH 7.4
• green= serial dilution was performed with a micropipet from A1,B1,C1,D1 - A11,B11,C11,D11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
• Measurement: FL, 530nm/30nm bandpass, 25-30nm with recommened excitation of 485nm, emission 520-530nm of the filter. Path length correction was turned off

Afternoon

LBC plates were made according to the protocol used on the wall

• 250ml LB 2x added to melted 250 ml WA 2x using a microwave
• 0.5ml was added to final solution
• plates were dried in 37C incubator

Transformation device 3 + negative control interlab study

• Device 3 (number 5) showed a low GFP expression, so it was tried to re-preform the tranformation. Negative control of the interlab (number 1) was not performed last time due to lack of LBC plates so was also performed.
• Protocol Transformation